US5117034A - Phosphatidylserine derivatives - Google Patents
Phosphatidylserine derivatives Download PDFInfo
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- US5117034A US5117034A US07/602,557 US60255790A US5117034A US 5117034 A US5117034 A US 5117034A US 60255790 A US60255790 A US 60255790A US 5117034 A US5117034 A US 5117034A
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- phosphatidylserine
- phosphatidylserine derivative
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- 150000008106 phosphatidylserines Chemical class 0.000 title claims abstract description 24
- 239000000203 mixture Substances 0.000 claims abstract description 11
- RUPAXCPQAAOIPB-UHFFFAOYSA-N tert-butyl formate Chemical group CC(C)(C)OC=O RUPAXCPQAAOIPB-UHFFFAOYSA-N 0.000 claims abstract description 11
- 125000004432 carbon atom Chemical group C* 0.000 claims abstract description 10
- 125000001424 substituent group Chemical group 0.000 claims abstract description 10
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 5
- 125000004453 alkoxycarbonyl group Chemical group 0.000 claims abstract description 5
- 125000004448 alkyl carbonyl group Chemical group 0.000 claims abstract description 5
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 5
- 239000002253 acid Substances 0.000 claims abstract description 4
- 125000000217 alkyl group Chemical group 0.000 claims abstract 3
- 125000002015 acyclic group Chemical group 0.000 claims description 15
- 125000001183 hydrocarbyl group Chemical group 0.000 claims 14
- 125000003118 aryl group Chemical group 0.000 abstract description 2
- 125000005843 halogen group Chemical group 0.000 abstract description 2
- 125000006239 protecting group Chemical group 0.000 abstract description 2
- 101100386054 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) CYS3 gene Proteins 0.000 abstract 1
- 101150035983 str1 gene Proteins 0.000 abstract 1
- 150000001875 compounds Chemical class 0.000 description 39
- 239000012528 membrane Substances 0.000 description 24
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 21
- 150000002632 lipids Chemical class 0.000 description 19
- 238000000034 method Methods 0.000 description 17
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 12
- 239000000463 material Substances 0.000 description 12
- 238000003786 synthesis reaction Methods 0.000 description 10
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 230000015572 biosynthetic process Effects 0.000 description 8
- 239000011541 reaction mixture Substances 0.000 description 8
- 239000002502 liposome Substances 0.000 description 7
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 6
- 125000003277 amino group Chemical group 0.000 description 6
- 238000000921 elemental analysis Methods 0.000 description 6
- 230000002829 reductive effect Effects 0.000 description 6
- 239000002904 solvent Substances 0.000 description 6
- CZDYPVPMEAXLPK-UHFFFAOYSA-N tetramethylsilane Chemical compound C[Si](C)(C)C CZDYPVPMEAXLPK-UHFFFAOYSA-N 0.000 description 6
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 5
- 125000000524 functional group Chemical group 0.000 description 5
- 239000010421 standard material Substances 0.000 description 5
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000005481 NMR spectroscopy Methods 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical group OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 3
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- DTQVDTLACAAQTR-UHFFFAOYSA-M Trifluoroacetate Chemical compound [O-]C(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-M 0.000 description 3
- 238000000862 absorption spectrum Methods 0.000 description 3
- 239000013543 active substance Substances 0.000 description 3
- 230000002411 adverse Effects 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 239000003431 cross linking reagent Substances 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 3
- 125000003607 serino group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(O[H])([H])[H] 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- MCTWTZJPVLRJOU-UHFFFAOYSA-N 1-methyl-1H-imidazole Chemical compound CN1C=CN=C1 MCTWTZJPVLRJOU-UHFFFAOYSA-N 0.000 description 2
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 229930186217 Glycolipid Natural products 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- -1 N-succinimidyl-3-(2-pyridyldithio)propionyl phosphatidylethanolamine Chemical class 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- BQADRZHPZVQGCW-LBPRGKRZSA-N benzyl (2s)-3-hydroxy-2-[(2-methylpropan-2-yl)oxycarbonylamino]propanoate Chemical compound CC(C)(C)OC(=O)N[C@@H](CO)C(=O)OCC1=CC=CC=C1 BQADRZHPZVQGCW-LBPRGKRZSA-N 0.000 description 2
- TXFOLHZMICYNRM-UHFFFAOYSA-N dichlorophosphoryloxybenzene Chemical compound ClP(Cl)(=O)OC1=CC=CC=C1 TXFOLHZMICYNRM-UHFFFAOYSA-N 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 238000004992 fast atom bombardment mass spectroscopy Methods 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N glycerol Substances OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 239000010410 layer Substances 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- UKVIEHSSVKSQBA-UHFFFAOYSA-N methane;palladium Chemical compound C.[Pd] UKVIEHSSVKSQBA-UHFFFAOYSA-N 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 238000010898 silica gel chromatography Methods 0.000 description 2
- 229910052938 sodium sulfate Inorganic materials 0.000 description 2
- 235000011152 sodium sulphate Nutrition 0.000 description 2
- JWDFQMWEFLOOED-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 3-(pyridin-2-yldisulfanyl)propanoate Chemical compound O=C1CCC(=O)N1OC(=O)CCSSC1=CC=CC=N1 JWDFQMWEFLOOED-UHFFFAOYSA-N 0.000 description 1
- HNTGIJLWHDPAFN-UHFFFAOYSA-N 1-bromohexadecane Chemical compound CCCCCCCCCCCCCCCCBr HNTGIJLWHDPAFN-UHFFFAOYSA-N 0.000 description 1
- LWCIBYRXSHRIAP-UHFFFAOYSA-N 3-phenylmethoxypropane-1,2-diol Chemical compound OCC(O)COCC1=CC=CC=C1 LWCIBYRXSHRIAP-UHFFFAOYSA-N 0.000 description 1
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 description 1
- 238000006845 Michael addition reaction Methods 0.000 description 1
- 101150108015 STR6 gene Proteins 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- AGEZXYOZHKGVCM-UHFFFAOYSA-N benzyl bromide Chemical compound BrCC1=CC=CC=C1 AGEZXYOZHKGVCM-UHFFFAOYSA-N 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 150000002430 hydrocarbons Chemical group 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000000269 nucleophilic effect Effects 0.000 description 1
- 239000012044 organic layer Substances 0.000 description 1
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
- C07F9/02—Phosphorus compounds
- C07F9/06—Phosphorus compounds without P—C bonds
- C07F9/08—Esters of oxyacids of phosphorus
- C07F9/09—Esters of phosphoric acids
- C07F9/12—Esters of phosphoric acids with hydroxyaryl compounds
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
- C07F9/02—Phosphorus compounds
- C07F9/06—Phosphorus compounds without P—C bonds
- C07F9/08—Esters of oxyacids of phosphorus
- C07F9/09—Esters of phosphoric acids
- C07F9/091—Esters of phosphoric acids with hydroxyalkyl compounds with further substituents on alkyl
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Definitions
- This invention relates to a phosphatidylserine derivative useful as a precursor for a lipid membrane structural material for modifying the surface of a lipid bimolecular membrane such as a liposome.
- lipid bimolecular membrane such as a liposome
- surface reformation modification
- Known methods of introducing a functional compound like a protein to a lipid bimolecular membrane such as a liposome involve using a non-covalent bond or using a covalent bond. Considering the strength of the bond and the stability of the membrane, among other things, the covalent bond method is more useful.
- the method of using a covalent bond involves combining the surface of a lipid bimolecular membrane with a chosen molecule for forming a covalent bond using a divalent crosslinking agent and practically, an amino group or an SH group on a protein, or a sugar chain is utilized as a functional group for the bonding.
- a method of using an SH group involves a method of bonding using a maleinimido group as a crosslinking agent via a Michael addition reaction for the maleinimido group (Biochimica et Biophysica Acta, 943, 53 (1988)), a method of bonding an antibody activated by N-succinimidyl-3-(2-pyridyldithio)propionyl phosphatidylethanolamine and N-succinimidyl-3-(2-pyridyldithio) propionate (Nature, 288, 602 (1980)), and a method of bonding using an ⁇ -haloketone group as the crosslinking agent via a nucleophilic displacement reaction for the group (European Patent 0,312,212).
- a method of utilizing sugar chains involves a method of incorporating a glycolipid into the membrane side of the lipid bimolecular membrane, cutting the glycolipid with periodic acid, and reacting the aldehyde formed with an amino group (Science, 210, 539 (1980)).
- JP-A lipid bimolecular membrane
- JP-A means an "unexamined published Japanese patent application”
- a single-strand lipid membrane structural material is a reverse corn type molecule wherein the hydrophilic portion is more bulky than the hydrophobic portion, it has a disadvantage of being easily released from the membrane component in which case it would become a membrane poison.
- lipid membrane structural material that can be easily used to modify the surface of a lipid bimolecular membrane, has a variety of applications, and is safe for living organisms.
- An object of this invention is to provide a phosphatidylserine derivative which can easily modify a surface of a lipid bimolecular membrane and is safe for living organisms.
- a phosphatidylserine derivative represented by formulae (I), (II), or (III): ##STR2## wherein R represents a protective group capable of being removed by an acid, (preferably a tertiary butoxycarbonyl group); R' represents a straight chain or branched acyclic hydrocarbon group having from 8 to 24 carbon atoms, (preferably having 14, 16 or 18 carbon atoms), and this group may have a substituent group or may be unsaturated. Examples of substituents are an alkylcarbonyl group, an alkoxycarbonyl group, a halogen atom, and an aryl group. If unsaturated, there may be a double bond or a triple bond, and there may be two or more such bonds in the same chain. If there is an asymmetric carbon in the molecule the derivative may be a racemic mixture or an optically active isomer thereof.
- FIG. 1 is a graph of the infrared absorption spectrum, using a nujol paste method, of an optically active substance of a compound corresponding to formula (I) wherein R is a tertiary butoxycarbonyl group and R' is n--C 16 H 33 .
- FIG. 2 is a graph of the 200 MHz proton nuclear magnetic resonance spectrum (solvent: CDCl 3 , standard material: tetramethylsilane) of the compound analyzed in FIG. 1.
- FIG. 3 is a graph of the infrared absorption spectrum, using a nujol paste method, of an optically active substance of a compound corresponding to formula (II) wherein R is a tertiary butoxycarbonyl group and R' is n--C 16 H 33 .
- FIG. 4 is a graph of the 200 MHz proton nuclear magnetic resonance spectrum (solvent: CDCl 3 , standard material: tetramethylsilane) of the compound analyzed in FIG. 3.
- FIG. 5 is a graph of the infrared absorption spectrum of the trifluoroacetate of an optically active substance corresponding to formula (III) wherein R' is n--C 16 H 33 .
- FIG. 6 is a graph of the 200 MHz proton nuclear magnetic resonance spectrum (solvent: CDCl 3 , standard material: tetramethylsilane) of the compound analyzed in FIG. 5.
- amino group, the carboxy group, and the phosphoric acid group of the serine moiety of the compounds of the invention corresponding to formula (I) can be separately protected and also can be selectively deprotected without adversely affecting other functional groups.
- lipid bimolecular membrane such as a liposome by themselves or by mixing them with other lipid membrane structural materials.
- Such compounds can have a positive, negative, or amphoteric charge.
- serine which is one of the constitutional components of the compound, is a naturally existing amino acid, these compounds are safe for living organisms.
- lipid membrane structural material having functional properties can be obtained.
- the lipid membrane structure material of the invention can form a liquid bimolecular membrane such as a liposome by itself or when mixed with other types of lipid membrane structural material. Additionally, a surface modification or strengthening of the structure can be achieved in such a lipid bimolecular membrane.
- saccharide existing at the surface of a cell membrane functions in intercellular information transfer, for example, and there is a possibility that a liposome can be targeted to a specific cell by the surface modification with saccharide.
- the compounds of this invention are important as precursors for functional lipid membrane structural materials.
- this ester was obtained at a yield of 80% from (s)-N-t-butyoxycarbonylserine and benzyl bromide.
- the product was dissolved in 250 ml of ethyl acetate and after adding 1.5 g of 10% palladium-carbon, the mixture was reacted for 8 hours in a hydrogen atmosphere.
- the reaction mixture obtained was poured into 100 ml of water and extracted 4 times with 100 ml of chloroform.
- the organic layers (the extracts) were combined with each other, washed once with 150 ml of water, and dried with anhydrous sodium sulfate.
- the sodium sulfate was then removed by filtration and the filtrate was concentrated under reduced pressure to provide a colorless oily product.
- IR ⁇ max (Nujol): 3260(m), 2930(s), 2860(s), 1745(s), 1705(s), 1600(m), 1495(m), 1270(s), 1210(s), 1165(s), 1065(s), and 1030(s) cm -1 .
- IR ⁇ max (Nujol): 3600-3000(br m), 1725(s), 1600(m), 1500(s), 1255(s), 1210(s), 1170(s), 1060(s), 1030(s), and 960(s) cm -1 .
- reaction mixture was concentrated under reduced pressure to provide an oily product which was dried under reduced pressure to provide 1.05 g (quantitative) of the desired compound as a colorless waxy material.
- IR ⁇ max (Nujol): 3600-3000(br), 2740-2200(br), 1765(s), 1675(s), 1600(m), 1500(m), 1215(s) , 1170(s), and 1.040 cm -1 .
Abstract
A phosphatidylserine derivative represented by formulae (I), (II), and (III): ##STR1## wherein R represents a protective group which can be removed by an acid; and R' represents a straight chain or branched alkyl group having from 8 to 24 carbon atoms and may contain a substituent group, or may be unsaturated, provided that if an asymmetric carbon is present, the phosphatidylserine derivative is a racemic mixture or an optically active isomer thereof. Preferably, R represents a tertiary butoxycarbonyl group; R' represents a straight chain or branched alkyl group having 14, 16, or 18 carbon atoms; and the substituent is an alkylcarbonyl group, an alkoxycarbonyl group, a halogen atom, or an aryl group.
Description
This invention relates to a phosphatidylserine derivative useful as a precursor for a lipid membrane structural material for modifying the surface of a lipid bimolecular membrane such as a liposome.
In order to strengthen the structure and modify the function of a lipid bimolecular membrane such as a liposome, surface reformation (modification) is an important consideration. Known methods of introducing a functional compound like a protein to a lipid bimolecular membrane such as a liposome involve using a non-covalent bond or using a covalent bond. Considering the strength of the bond and the stability of the membrane, among other things, the covalent bond method is more useful.
Generally, the method of using a covalent bond involves combining the surface of a lipid bimolecular membrane with a chosen molecule for forming a covalent bond using a divalent crosslinking agent and practically, an amino group or an SH group on a protein, or a sugar chain is utilized as a functional group for the bonding.
More specifically, a method of using an SH group involves a method of bonding using a maleinimido group as a crosslinking agent via a Michael addition reaction for the maleinimido group (Biochimica et Biophysica Acta, 943, 53 (1988)), a method of bonding an antibody activated by N-succinimidyl-3-(2-pyridyldithio)propionyl phosphatidylethanolamine and N-succinimidyl-3-(2-pyridyldithio) propionate (Nature, 288, 602 (1980)), and a method of bonding using an α-haloketone group as the crosslinking agent via a nucleophilic displacement reaction for the group (European Patent 0,312,212). A method of utilizing sugar chains involves a method of incorporating a glycolipid into the membrane side of the lipid bimolecular membrane, cutting the glycolipid with periodic acid, and reacting the aldehyde formed with an amino group (Science, 210, 539 (1980)).
However, these methods all require a considerable number of steps and they can only be used for specific reactions.
It is also known to add a functional group to a lipid bimolecular membrane such as a liposome using a single-strand lipid membrane structural material which can be easily synthesized, as disclosed in JP-A-61-112021, JP-A-62-201864, JP-A-62-209092, and JP-A-1-27637 (the term "JP-A" as used herein means an "unexamined published Japanese patent application"). However, since a single-strand lipid membrane structural material is a reverse corn type molecule wherein the hydrophilic portion is more bulky than the hydrophobic portion, it has a disadvantage of being easily released from the membrane component in which case it would become a membrane poison.
Thus, there is great demand for a lipid membrane structural material that can be easily used to modify the surface of a lipid bimolecular membrane, has a variety of applications, and is safe for living organisms.
An object of this invention is to provide a phosphatidylserine derivative which can easily modify a surface of a lipid bimolecular membrane and is safe for living organisms.
It has been discovered that this and other objects can be attained by a phosphatidylserine derivative represented by formulae (I), (II), or (III): ##STR2## wherein R represents a protective group capable of being removed by an acid, (preferably a tertiary butoxycarbonyl group); R' represents a straight chain or branched acyclic hydrocarbon group having from 8 to 24 carbon atoms, (preferably having 14, 16 or 18 carbon atoms), and this group may have a substituent group or may be unsaturated. Examples of substituents are an alkylcarbonyl group, an alkoxycarbonyl group, a halogen atom, and an aryl group. If unsaturated, there may be a double bond or a triple bond, and there may be two or more such bonds in the same chain. If there is an asymmetric carbon in the molecule the derivative may be a racemic mixture or an optically active isomer thereof.
FIG. 1 is a graph of the infrared absorption spectrum, using a nujol paste method, of an optically active substance of a compound corresponding to formula (I) wherein R is a tertiary butoxycarbonyl group and R' is n--C16 H33.
FIG. 2 is a graph of the 200 MHz proton nuclear magnetic resonance spectrum (solvent: CDCl3, standard material: tetramethylsilane) of the compound analyzed in FIG. 1.
FIG. 3 is a graph of the infrared absorption spectrum, using a nujol paste method, of an optically active substance of a compound corresponding to formula (II) wherein R is a tertiary butoxycarbonyl group and R' is n--C16 H33.
FIG. 4 is a graph of the 200 MHz proton nuclear magnetic resonance spectrum (solvent: CDCl3, standard material: tetramethylsilane) of the compound analyzed in FIG. 3.
FIG. 5 is a graph of the infrared absorption spectrum of the trifluoroacetate of an optically active substance corresponding to formula (III) wherein R' is n--C16 H33.
FIG. 6 is a graph of the 200 MHz proton nuclear magnetic resonance spectrum (solvent: CDCl3, standard material: tetramethylsilane) of the compound analyzed in FIG. 5.
Examples of the syntheses of optically active compounds according to the invention are illustrated below, but these syntheses are not limiting. ##STR3##
The amino group, the carboxy group, and the phosphoric acid group of the serine moiety of the compounds of the invention corresponding to formula (I) can be separately protected and also can be selectively deprotected without adversely affecting other functional groups.
When the carboxy group of the serine moiety of the compounds of the invention corresponding to formula (II) is in a free state, the amino and phosphoric acid groups thereof can be separately protected and also selectively deprotected without adversely affecting other functional groups.
When the amino group of the serine moiety of the compounds of this invention corresponding to formula (III) is in a state of high reactivity, the carboxy and phosphoric acid groups thereof can be separately protected and also selectively deprotected without adversely affecting other functional groups.
Compounds obtained by deprotecting a compound corresponding to formulae (I), (II), or (III) can form a lipid bimolecular membrane such as a liposome by themselves or by mixing them with other lipid membrane structural materials. Such compounds can have a positive, negative, or amphoteric charge. In addition, since serine, which is one of the constitutional components of the compound, is a naturally existing amino acid, these compounds are safe for living organisms.
Also, by bonding a functional compound such as an amino acid, peptide, protein, saccharide, polymerizable group, or spacer having a reactivity to one or both of the hydrophilic amino group or the carboxy group as a foothold, a lipid membrane structural material having functional properties can be obtained. The lipid membrane structure material of the invention can form a liquid bimolecular membrane such as a liposome by itself or when mixed with other types of lipid membrane structural material. Additionally, a surface modification or strengthening of the structure can be achieved in such a lipid bimolecular membrane.
It has recently been clarified that the saccharide existing at the surface of a cell membrane functions in intercellular information transfer, for example, and there is a possibility that a liposome can be targeted to a specific cell by the surface modification with saccharide.
Thus, the compounds of this invention are important as precursors for functional lipid membrane structural materials.
The invention is illustrated further with the non-limiting examples below. Unless otherwise indicated, all ratios and percentages are by weight.
Synthesis of an optically active compound of formula (I) when R is a tertiary butoxycarbonyl group and R' is n--C16 H33 :
A) Synthesis of (s)-N-t-Butoxycarbonylserine benzyl ester: ##STR4##
According to the method described in Synthesis, 961 (1979), this ester was obtained at a yield of 80% from (s)-N-t-butyoxycarbonylserine and benzyl bromide.
B) Synthesis of (s)-2,3-di-o-hexadecyl-1-glycerol: ##STR5##
In 300 ml of toluene was dissolved 12.0 g of glycerol monobenzyl ether prepared by the method described in Synthesis, 503 (1985). After adding 16.0 g of powdery potassium hydroxide and 84.0 g of hexadecyl bromide to the solution, the reaction mixture was refluxed under heating for 8 hours. The reaction mixture was allowed to cool to room temperature and diluted with 400 ml of hexane. The reaction mixture was then washed twice with 200 ml of water and dried with anhydrous sodium sulfate. After removing the sodium sulfate by filtration, the filtrate was concentrated under reduced pressure to provide a colorless oily product.
The reaction mixture was purified by silica gel chromatography (eluent: hexane/ethyl acetate=40/1) to provide 41.2 g (yield 95.5%) of a dialkylglycerol monobenzyl ether compound.
The properties of the product coincided with described in Biochemistry, 2, 394 (1963).
The product was dissolved in 250 ml of ethyl acetate and after adding 1.5 g of 10% palladium-carbon, the mixture was reacted for 8 hours in a hydrogen atmosphere.
Insoluble matter was removed using sellite filtration and the sellite layer was washed with ethyl acetate. The filtrate was combined with the washed solution and the mixture was concentrated under reduced pressure. The residue thus formed was recrystallized from ethyl acetate to provide the desired compound as colorless crystals.
The properties of the product coincided with those described in Biochemistry, 2, 394 (1963).
C) Synthesis of an optically active compound of formula (I) when R is a tertiary butoxycarbonyl group and R' is n--C16 H33 : ##STR6##
To a dry tetrahydrofuran solution of phenylphosphoro dichloridate (PhOPOCl2, 2.75 g, commercially available product) was added 20 ml of a dry tetrahydrofuran solution of 2.95 g of (s)-N-t-butoxycarbonylserine benzyl ester and 1.07 g of N-methylimidazole over a period of 20 minutes. After stirring the reaction mixture for 10 minutes at room temperature, 20 ml of a dry tetrahydrofuran solution of 5.4 g of (s)-2,3-di-o-hexadecyl-1-glycerol and 1.07 g of N-methylimidazole was added thereto over a period of 10 minutes and the mixture was allowed to stand for 14 hours at room temperature.
The reaction mixture obtained was poured into 100 ml of water and extracted 4 times with 100 ml of chloroform. The organic layers (the extracts) were combined with each other, washed once with 150 ml of water, and dried with anhydrous sodium sulfate. The sodium sulfate was then removed by filtration and the filtrate was concentrated under reduced pressure to provide a colorless oily product.
The reaction mixture obtained was purified by silica gel chromatography (eluent: hexane/ethyl acetate =20/1 to 8/1) to provide 5.48 g (yield 56.3%) of the desired compound as a colorless waxy material.
The properties of the product are shown below.
IR νmax (Nujol): 3260(m), 2930(s), 2860(s), 1745(s), 1705(s), 1600(m), 1495(m), 1270(s), 1210(s), 1165(s), 1065(s), and 1030(s) cm-1.
1 H NMR δ(200 MHz, solvent CDCl3, standard substance TMS):
0.87(6H, deformed t, J=6 Hz), 1.25(54H, br s), 1.45(9H, s), 1.43-1.60(4H, br), 3.36-3.66(7H, m), 4.05-4.30(2H, m), 4.33-4.65(3H, m), 5.08-5.24(2H, m), 5.48-5.68(1H, m), 7.11-7.40(10H, m), and FAB-MS 974[(M+H)+ ].
Following the method described in Example 1, except for the starting materials used, an optically active compound of formula (I) wherein R was a tertiary butoxycarbonyl group and R' was n--C14 H29 or n--C18 H37 was synthesized.
The properties of this compound are shown below.
A compound of formula (I) wherein R' was n--C14 H29 :
Elemental Analysis:
______________________________________
C H N
______________________________________
Found: 68.31% 9.61% 1.50%
Calculated:
68.05% 9.60% 1.53%
______________________________________
A compound of formula (I) wherein R' was n--C18 H37 :
______________________________________
C H N
______________________________________
Found: 69.86% 10.06% 1.41%
Calculated:
69.97% 10.11% 1.36%
______________________________________
Synthesis of an optically active compound of formula (II) wherein R is a tertiary butoxycarbonyl group and R' is n--C16 H33 :
In 10 ml of a mixed solvent of ethyl acetate and methanol (1:1) was dissolved 200 mg of compound obtained in C) of Example 1 and after adding thereto 10 mg of 5% palladium-carbon, the mixture was reacted for 7 hours at normal pressure in a hydrogen atmosphere.
Insoluble matter was removed by sellite filtration and the sellite layer was washed with ethyl acetate. The filtrate was combined with the washed solution and the mixture was concentrated under reduced pressure to provide 190 mg (quantitative) of the desired compound as a colorless waxy material.
The properties of the compound are shown below.
IR νmax (Nujol): 3600-3000(br m), 1725(s), 1600(m), 1500(s), 1255(s), 1210(s), 1170(s), 1060(s), 1030(s), and 960(s) cm-1.
1 H NMR δ(200 MHz, solvent CDCl3, standard material TMS):
0.88(6H, deformed t, J=6 Hz), 1.25(54H, br s), 1.43 and 1.66(9H, each s), 1.40-1.60(4H, m), 3.38-3.66(7H, m), 4.07-4.65(5H, m), 5.60-5.70(1H, m), 6.58-7.00(1H, br, COOH), 7.10-7.40(5H, m), and FAB-MS 906 [(M+Na)+ ].
Following the method described in Example 3, except for the starting materials used, an optically active compound of formula (II) wherein R was a tertiary butoxycarbonyl group and R' was n--C14 H29 or n--C18 H37 was synthesized.
The properties of this compound are shown below.
A compound of formula (II) wherein R' was n--C14 H29 :
Elemental Analysis:
______________________________________
C H N
______________________________________
Found: 65.26% 9.94% 1.63%
Calculated:
65.30% 9.92% 1.69%
______________________________________
A compound of formula (II) wherein R' was n--C18 H37 :
Elemental Analysis:
______________________________________
C H N
______________________________________
Found: 69.56% 10.41% 1.37%
Calculated:
67.73% 10.44% 1.49%
______________________________________
Synthesis of the trifluoroacetate of an optically active compound of formula (III) wherein R' is n--C16 H33 :
After adding 10 ml of trifluoroacetic acid (commercially available product) to 10 ml of a methylene chloride solution of 1.0 g of compound obtained in C) of Example 1, the mixture was stirred for 20 minutes at room temperature.
The reaction mixture was concentrated under reduced pressure to provide an oily product which was dried under reduced pressure to provide 1.05 g (quantitative) of the desired compound as a colorless waxy material.
The properties thereof are shown below.
IR νmax (Nujol): 3600-3000(br), 2740-2200(br), 1765(s), 1675(s), 1600(m), 1500(m), 1215(s) , 1170(s), and 1.040 cm-1.
1 H NMR δ(200 MHz, solvent CDCl3, standard material TMS):
0.87(6H, deformed t, J=6 Hz), 1.25(54H, br s), 1.40-1.60(4H, m), 3.35-3.63(7H, m), 4.00-4.30(2H, m), 4.40-4.50(1H, m), 4.59-4.64(2H, m), 5.13-5.21(2H, m), 7.07-7.43(10H, m), and 7.80-8.20(3H, amino group exchanging proton).
Elemental Analysis:
______________________________________
C H N
______________________________________
Found: 64.21% 9.18% 1.41%
Calculated:
64.43% 9.02% 1.42%
______________________________________
Following the method described in Example 5, except for the starting materials used, a trifluoroacetate of an optically active compound of formula (III) wherein R' was n--C14 H29 or n--C18 H37 was synthesized.
The properties thereof are shown below.
Compound of formula (III) wherein R' was n--C14 H29 :
Elemental Analysis:
______________________________________
C H N
______________________________________
Found: 62.99% 8.61% 1.43%
Calculated:
63.16% 8.70% 1.50%
______________________________________
Compound of formula (III) wherein R' was n--C18 H37 :
Elemental Analysis:
______________________________________
C H N
______________________________________
Found: 65.53% 9.15% 1.42%
Calculated:
65.58% 9.30% 1.34%
______________________________________
While the invention has been described in detail and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof.
Claims (15)
1. A phosphatidylserine derivative represented by formula (I): ##STR7## wherein R represents a tertiary butoxycarbonyl group which can be removed with an acid; and R' represents a straight chain or branched acyclic hydrocarbon group having from 8 to 24 carbon atoms, and may have a substituent group, wherein said substituent group is an alkylcarbonyl group or alkoxycarbonyl group, or may be unsaturated, provided that if an asymmetric carbon is present, said phosphatidylserine derivative is a racemic mixture or an optically active isomer thereof.
2. A phosphatidylserine derivative represented by formula (II): ##STR8## wherein R represents a tertiary butoxycarbonyl group which can be removed with an acid; and R' represents a straight chain or branched acyclic hydrocarbon group having from 8 to 24 carbon atoms, and may have a substituent group, wherein said substituent group is an alkylcarbonyl group or alkoxycarbonyl group, or may be unsaturated, provided that if an asymmetric carbon is present, said phosphatidylserine derivative is a racemic mixture or an optically active isomer thereof.
3. A phosphatidylserine derivative represented by formula (III): ##STR9## wherein R' represents a straight chain or branched acyclic hydrocarbon group having from 8 to 24 carbon atoms, and may have a substituent group, wherein said substituent group is an alkylcarbonyl group or alkoxycarbonyl group, or may be unsaturated, provided that if an asymmetric carbon is present, said phosphatidylserine derivative is a racemic mixture or an optically active isomer thereof.
4. A phosphatidylserine derivative as claimed in claim 1, wherein R' represents a straight chain or branched alkyl group having 14, 16, or 18 carbon atoms.
5. A phosphatidylserine derivative as claimed in claim 1, wherein said acyclic hydrocarbon group contains at least one double bond.
6. A phosphatidylserine derivative as claimed in claim 1, wherein said acyclic hydrocarbon group contains at least one triple bond.
7. A phosphatidylserine derivative as claimed in claim 1, wherein said acyclic hydrocarbon group contains a combination of double and triple bonds.
8. A phosphatidylserine derivative as claimed in claim 2, wherein R' represents a straight chain or branched acyclic hydrocarbon group having 14, 16, or 18 carbon atoms.
9. A phosphatidylserine derivative as claimed in claim 2, wherein said acyclic hydrocarbon group contains at least one double bond.
10. A phosphatidylserine derivative as claimed in claim 2, wherein said acyclic hydrocarbon group contains at least one triple bond.
11. A phosphatidylserine derivative as claimed in claim 2, wherein said acyclic hydrocarbon group contains a combination of double and triple bonds.
12. A phosphatidylserine derivative as claimed in claim 3, wherein R' represents a straight chain or branched acyclic hydrocarbon group having 14, 16, or 18 carbon atoms.
13. A phosphatidylserine derivative as claimed in claim 3, wherein said acyclic hydrocarbon group contains at least one double bond.
14. A phosphatidylserine derivative as claimed in claim 3, wherein said acyclic hydrocarbon group contains at least one triple bond.
15. A phosphatidylserine derivative as claimed in claim 3, wherein said acyclic hydrocarbon group contains a combination of double and triple bonds.
Applications Claiming Priority (8)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1-276525 | 1989-10-24 | ||
| JP27652589 | 1989-10-24 | ||
| JP1-278014 | 1989-10-25 | ||
| JP1-278013 | 1989-10-25 | ||
| JP27801389 | 1989-10-25 | ||
| JP27801489 | 1989-10-25 | ||
| JP2137887A JPH03204889A (en) | 1989-10-24 | 1990-05-28 | Phosphatidylserine derivative |
| JP2-137887 | 1990-05-28 |
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| Publication Number | Publication Date |
|---|---|
| US5117034A true US5117034A (en) | 1992-05-26 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| US07/602,557 Expired - Lifetime US5117034A (en) | 1989-10-24 | 1990-10-24 | Phosphatidylserine derivatives |
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| Country | Link |
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| US (1) | US5117034A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5965413A (en) * | 1995-11-08 | 1999-10-12 | Kabushiki Kaisha Yakult Honsha | Process for producing phosphatidylserines having long chain unsaturated fatty acid as side chain |
| US20030147944A1 (en) * | 1999-12-10 | 2003-08-07 | Mayer Lawrence D | Lipid carrier compositions with protected surface reactive functions |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1989010370A1 (en) * | 1988-04-19 | 1989-11-02 | Cl-Pharma Aktiengesellschaft | New alkylphosphono- and phosphoserines, method for preparing them, and pharmaceutical substances containing them |
-
1990
- 1990-10-24 US US07/602,557 patent/US5117034A/en not_active Expired - Lifetime
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1989010370A1 (en) * | 1988-04-19 | 1989-11-02 | Cl-Pharma Aktiengesellschaft | New alkylphosphono- and phosphoserines, method for preparing them, and pharmaceutical substances containing them |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5965413A (en) * | 1995-11-08 | 1999-10-12 | Kabushiki Kaisha Yakult Honsha | Process for producing phosphatidylserines having long chain unsaturated fatty acid as side chain |
| US20030147944A1 (en) * | 1999-12-10 | 2003-08-07 | Mayer Lawrence D | Lipid carrier compositions with protected surface reactive functions |
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